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Part:BBa_K1119009:Design

Designed by: Hyun Jung Lee   Group: iGEM13_Hong_Kong_HKUST   (2013-09-16)

CMV promoter - MLS - GFP - hGH polyA tail


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 614
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 628
    Illegal AgeI site found at 1462
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1865
    Illegal BsaI.rc site found at 1367


Design Notes

There are NgoMIV & AgeI sites around MLS CDS (BBa_K1119001) and GFP reporter CDS(BBa_K648013), which make the part incompatible with RFC25 standard, but users can perform in-frame protein fusion using the mentioned sites.

Source

The CMV promoter (BBa_K1119006) sequence was cloned out from pEGFP-N1(Clonetech)using PCR with primers that includes prefix and suffix in RFC10 standard.

MLS sequence in RFC25 format from partsregistry (BBa_K1119001)

GFP reporter in RFC25 format from partsregistry(BBa_K648013)

hGH polyA terminator in RFC10 format from partsregistry (BBa_K404108)

References

Clonetech.(1999).pEGFP-N1 Vector Information.Retrieved from http://www.pkclab.org/PKC/vector/pEGFPN1.pdf